screening
models of
Alzheimer’s
disease
Experimental Pharmaacology
Guided by Dr Hari Prasad MG
Presented by
Moksha Ponnamma PG
Om Balaji Biradar
Shubham M Hosamani
Sneha A Bhat
Sonu M A
What is Alzheimer’s?
It is a neurodegenerative disease of the cerebral cortex
with decreased cholinergic transmission and is the most
common cause of dementia (loss of memory, reduced
thinking, behavioral & social skills).
It was 1st described Dr. Alois Alzheimer, German physician
in 1906.
It is caused by several genetical & environmental factors.
Other factors include :
1. Advancing age
2. Family history
3. Trauma
4. Vascular disease like stroke
Risk Factors
01 02 03
Increased
Increase Chloesterol
d age Hypertension levels
04 05 06
Coronary Smoking and
Arterty disease Diabetes Alcohol use
Symptoms
Pathophysiology
Alzheimer’s disease is associated with brain shrinkage &
localized loss of neurons, mainly in the hippocampus & basal
forebrain. The loss of cholinergic neuron in the hippocampus &
frontal cortex is the feature of the disease & is thought to
underline the cognitive deficit & loss of short-term memory that
occur in AD.
Two microscopic feature are characteristic of the disease ,
namely extracellular amyloid plaques consisting of
extracellular deposits of β amyloid protein & intra neuronal
neurofibrillary tangles comprising filaments of a
phosphorylated form of a microtubule associated protein ( Tau).
Drugs used:
These are diseases that are These are long-term These are diseases that are
caused by pathogenic illnesses that typically caused by abnormal genes
microorganisms such as progress slowly and may or chromosomal
bacteria, viruses, fungi or not have a cure. Chronic abnormalities. Genetic
parasites. Examples of diseases can be caused by diseases can be inherited
infectious diseases include a variety of factors or they may occur
the flu or tuberculosis spontaneously
SCREENING METHODS
Passive avoidance Active avoidance
1. Step down 1. Shuttle box avoidance
2. Step through 2. Jumping avoidance
3. Two compartment test 3. Runway avoidance
4. Up hill avoidance
Discrimintaion
learning
1. Open field test
2. Radial arm maze
3. Morris water maze
4. Plus maze
STEP DOWN
An animal (mouse or rat) in an open field spends most of the time
close to the walls and in the corners. When placed on an elevated
platform in the center of a rectangular compartment, it steps down
almost immediately to the floor to explore the enclosure and to
approach the wall.
1. Familiarization
2. Learning
3. Retention Test
Evaluation: The time of descent during the learning phase
and the time during the retention test is measured. A
prolongation of the step-down latency is defined as learning.
Evaluation: The time to step- through during the learning phase is measured & the
time during the retention test is measured.
In this test a prolongation of the step through latencies is specific to the
experimental situation.
RADIAL ARM MAZE
• The radial arm maze allows the study of spatial reference and working memory
processes in the rat.
• The rat uses spatial information provided by the distal cues in the room to efficiently
locate the baited arms
• Food pellets are placed at the end of the arms .
• During the test , rats are fed once a day and their body weights maintained at 85% of
their free feeding weight to motivate the rat to run the maze
• Animals are trained on a daily basis in the maze to collect the food pellets
Evaluation : The number of errors (entries to non-baited arms ) are
counted during the session
MORRIS WATER
A task was developed where ratsMAZE
learn to swim in a water tank to find an escape platform hidden under the water .
The water tank is divided in 4 equal quadrants and a small platform is located in the center of one of the quadrants .
The rat is released into the water and allowed 60-90 s to find the platform
The platform remains in the same position during the training days
Animals usually receive 2 to 4 trails per days for 4 to 5 days until they escape onto the platform .Well trained rats
escape in 10 s
IN VITRO
METHODS
Inhibition of acetylcholine- esterase activity in rat
striatum
Long-term potentiation in hippocampal slices
Inhibition of acetylcholine- esterase activity in rat
striatum
TISSUE PREPERATION
Male Wistar rats are decapitated
Brains rapidly removed corpora striata dissected free
Weighed & homogenized in 19 volumes of 0.05 M NaH 2PO4
PH 7.2 using a Potter- Elvejhem homogenizer
A 25 µl aliquot of this suspension is added to 1ml of the vehicle or various
concentrations of the test drug
Re- incubated for 10 min at 37°c
Assay
Long-term potentiation in hippocampal slices
• It is an example of activity dependent synaptic plasticity that has
been identified in mammalian brain.
• A brief tetanus to any one of a number of monosynaptic excitatory
pathways in the hippocampus can enhance the amplitude of evoked
responses in the tetanized pathway for days thereafter.
• The extracellular population spike is obtained using glass microelectrodes filled with 2 M NaCl
which have resistances of 2-5 MΩ .
• The electrodes are placed into the stratum pyramidale of CA1 or CA3
• The signal is amplified and stored on magnetic discs for later analysis
• The magnitude of the population spike is evaluated by taking the voltage difference between
negative peak & the following positive peak.
● Constant current pulses or delivered with the frequency of 0.2Hz , only during
the test intervals .
● The stimulation intensity is adjusted to elicit the population spike 40% &80%
of its maximal amplitude in CA1 &CA3 are respectively .
● After the base line is recorded for 10 to 20 min , LTP is induced by repetitive
stimulation of 100 pulses at 20 Hz for 5s in CA1 & at 50Hz for 2s in CA3 at the
same strength as for the test pulses .
● Responses by test pulses are recorded 0 , 10 ,20 & 30min after repetitive
stimulation .
● The test drugs are dissolved in the cerebrospinal fluid and applied
extracellulary at various concentrations by switching perfusion reservoir
EVALUATION
● The time course of LTP is registered for CA1 & CA3.
● The mean percent increase in the amplitude of the population spike from
REFERENCE
1. Drug screening methods ( Preclinical Evaluation of New drugs) by
S.K Gupta 2nd Edition
2. Drug Discovery & Evaluation by H. Gerhard Vogel 2 nd Edition
3. Principles of pharmacology HL Sharma KK Sharma 2 nd Edition
4. Pharmacology H.P. Rang , M.M. Dale , J.M Riffer ,
P.K. Moore
Thanks
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